Hepatitis C virus diversity and treatment outcomes in Benin: a prospective cohort study.

BACKGROUND: 10 million people are chronically infected with the hepatitis C virus (HCV) in sub-Saharan Africa. The assessment of viral genotypes and treatment response in this region is necessary to achieve the WHO target of worldwide elimination of viral hepatitis by 2030. We aimed to investigate the prevalence of HCV genotypes and outcomes of treatment with direct-acting antiviral agents in Benin, a country with a national HCV seroprevalence of 4%. METHODS: This prospective cohort study was conducted at two referral hospitals in Benin. Individuals were eligible for inclusion if they were seropositive for HCV and willing to consent to participation in the study; exclusion criteria were an inability to give consent or incarceration. Viraemia was confirmed by PCR. The primary outcomes were to identify HCV genotypes and measure sustained virological response rates 12 weeks after completion of treatment (SVR12) with a 12-week course of sofosbuvir-velpatasvir or sofosbuvir-ledipasvir, with or without ribavirin. We conducted phylogenetic and resistance analyses after the next-generation sequencing of samples with a cycle threshold (Ct) value of 30 or fewer cycles. The in-vitro efficacy of NS5A inhibitors was tested using a subgenomic replicon assay. FINDINGS: Between June 2, 2019, and Dec 30, 2020, 148 individuals were screened for eligibility, of whom 100 were recruited prospectively to the study. Plasma samples from 79 (79%) of the 100 participants were positive for HCV by PCR. At the time of the study, 52 (66%) of 79 patients had completed treatment, with an SVR12 rate of 94% (49 of 52). 57 (72%) of 79 samples had a Ct value of 30 or fewer cycles and were suitable for whole-genome sequencing, from which we characterised 29 (51%) samples as genotype 1 and 28 (49%) as genotype 2. Three new genotype 1 subtypes (1q, 1r, and 1s) and one new genotype 2 subtype (2xa) were identified. The most commonly detected subtype was 2d (12 [21%] of 57 samples), followed by 1s (eight [14%]), 1r (five [9%]), 1b (four [7%]), 1q (three [5%]), 2xa (three [5%]), and 2b (two [3%]). 20 samples (11 genotype 2 and nine genotype 1) were unassigned new singleton lineages. 53 (93%) of 57 sequenced samples had at least two resistance-associated substitutions within the NS5A gene. Subtype 2d was associated with a lower-than-expected SVR12 rate (eight [80%] of ten patients). For one patient, with subtype 2b, treatment was not successful. INTERPRETATION: This study revealed a high SVR rate in Benin among individuals treated for HCV with sofosbuvir-velpatasvir, including those with highly diverse viral genotypes. Further studies of treatment effectiveness in genotypes 2d and 2b are indicated. FUNDING: Medical Research Council, Wellcome, Global Challenges Research Fund, Academy of Medical Sciences, and PHARMBIOTRAC.

Gaps and opportunities in sustainable medicines use in resource limited settings: A situational analysis of Uganda.

Resource constraints and widespread poverty among populations undermine disease prevention in low- and middle-income countries (LMICs) and ensure that these countries carry a disproportionate share of the global disease burden. Lack of access to efficacious medicines in LMICs further exacerbates this inequity. Addressing inequitable access to medicines and assuring their sustainable use is critical to the well-being of these populations. Whilst inadequate access to medicines in LMICs has drawn much attention, less is known about the sustainable use of available medicines, particularly to ensure their efficacy and mitigate harm to the population and the environment. Uganda has adopted various measures to ensure sustainable medicines use, including a national medicines policy, essential medicines list, medicines regulation framework and promotion of domestic medicines production. Despite progress, challenges remain to achieving sustainable medicines use in the country, including fragmented access, inappropriate use, poor quality and inappropriate disposal. There is a need to consolidate the globally embraced One Health approach (fostering collaboration between human, animal and environmental health sectors) to addressing these challenges as espoused in the country's One Health strategic plan. Medicines supply chain management in public sector health facilities needs to be strengthened to minimize inventory shortages (stock-outs). A strategy for universal health insurance can minimize economic barriers to medicines access. Enhanced professional and medicines regulation in the private health market needs to be implemented. There are opportunities to build further capacity in Uganda, particularly infrastructure for regulation of its healthcare systems, policy and governance, workforce capacity building, and population action and engagement.

The 27 kDa Trypanosoma brucei Pentatricopeptide Repeat Protein is a G-tract Specific RNA Binding Protein.

Pentatricopeptide repeat (PPR) proteins, a helical repeat family of organellar RNA binding proteins, play essential roles in post-transcriptional RNA processing. In Trypanosoma brucei, an expanded family of PPR proteins localize to the parasite's single mitochondrion, where they are believed to perform important roles in both RNA processing and translation. We studied the RNA binding specificity of the simplest T. brucei PPR protein (KRIPP11) using electrophoretic mobility shift assays, fluorescence anisotropy, circular dichroism spectroscopy, and in vitro selection. We found KRIPP11 to be an RNA binding protein with specificity for sequences of four or more consecutive guanosine residues (G-tracts). Such G-tracts are dramatically enriched in T. brucei mitochondrial transcripts that are destined for extensive uridine insertion/deletion editing but are not present in mRNAs following editing. We further found that the quadruplex oligoguanosine RNA conformation is preferentially recognized by KRIPP11 over other conformational forms, and is bound without disruption of the quadruplex structure. In combination with prior data demonstrating association of KRIPP11 with the small ribosomal subunit, these results suggest possible roles for KRIPP11 in bridging mRNA maturation and translation or in facilitating translation of unusual dual-coded open reading frames.

Participatory healthcare-provider orientation to improve artemether-lumefantrine-based drug treatment of uncomplicated malaria: a cluster quasi-experimental study.

OBJECTIVES: To assess the effect of participatory healthcare-provider orientation in enhancing patient knowledge, appropriate prescribing and dispensing of artemether-lumefantrine, during drug treatment of uncomplicated malaria. METHODS: A cluster quasi-experimental study. The authors developed strategies to address challenges encountered by healthcare providers during clinical management of malaria. The primary outcome was patient knowledge on prescribed malaria drug treatment. Secondary outcomes were appropriate prescribing and provision of adequate drug dispensing information. The authors used generalised estimating equation logistic regression to investigate correlates of appropriate use of artemether-lumefantrine. RESULTS: The proportion of patients or caretakers of paediatric patients sufficiently knowledgeable about malaria treatment increased from 16/85 (18.8%) at baseline to 33/96 (34.4%) at evaluation, OR 2.26 (95% CI 1.13 to 4.49), p=0.020, in the intervention, and fell slightly from 49/134 (36.6%) to 35/114 (30.7%), OR 0.77, (95% CI 0.45 to 1.31), p=0.331 in the control district. This was enhanced by the existence of drug-dispensing standard operating procedures (adjusted OR 1.85, 95% CI 0.98 to 3.50; p=0.057). The proportion of appropriate prescriptions increased from 61/87 (70.1%) to 94/112 (83.9%) in the intervention district, OR 2.23 (95% CI 1.13 to 4.40), p=0.020 and reduced from 91/115 (79.1%) to 75/112 (67.0%) in the control district, OR 0.53, (95% CI 0.29 to 0.97), p=0.040. The frequency of adequately dispensed prescriptions increased in the intervention district (34(32.4%) to 53(45.3%), OR 1.73 (95% CI 1.00 to 2.99), p=0.050) but decreased in the control location (94 (69.6%) to 71 (52.6%), OR 0.48 (95% CI 0.29 to 0.80), p=0.004). CONCLUSIONS: Participatory healthcare-provider orientation enhanced patient knowledge, healthcare provider prescribing and dispensing of artemether-lumefantrine, bolstered by adequate medication counselling and use of drug-dispensing standard operating procedures.